Producing multiple chemicals through biological upcycling of waste poly(ethylene terephthalate).

Poly(ethylene terephthalate) (PET) waste is of low degradability in nature, and its mismanagement threatens numerous ecosystems. To combat the accumulation of waste PET in the biosphere, PET bio-upcycling, which integrates chemical pretreatment to produce PET-derived monomers with their microbial conversion into value-added products, has shown promise. The recently discovered Rhodococcus jostii RPET strain can metabolically degrade terephthalic acid (TPA) and ethylene glycol (EG) as sole carbon sources, and it has been developed into a microbial chassis for PET upcycling. However, the scarcity of synthetic biology tools, specifically designed for this non-model microbe, limits the development of a microbial cell factory for expanding the repertoire of bioproducts from postconsumer PET. Herein, we describe the development of potent genetic tools for RPET, including two inducible and titratable expression systems for tunable gene expression, along with serine integrase-based recombinational tools (SIRT) for genome editing. Using these tools, we systematically engineered the RPET strain to ultimately establish microbial supply chains for producing multiple chemicals, including lycopene, lipids, and succinate, from postconsumer PET waste bottles, achieving the highest titer of lycopene ever reported thus far in RPET [i.e., 22.6 mg/l of lycopene, ~10 000-fold higher than that of the wild-type (WT) strain]. This work highlights the great potential of plastic upcycling as a generalizable means of sustainable production of diverse chemicals.